This R21 application is developed in response to our recent findings, which indicate that women treated by loop electrosurgical excision procedure (LEEP) for surgical removal of cervical intra-epithelial neoplasia (CIN) face a two-fold increased risk of preterm delivery due to preterm premature rupture of the membranes (pPROM). We hypothesize that LEEP may alter the cervix in regard to its structure, anatomy, and composition of cervical secretions. An adverse effect of LEEP might operate by reducing cervical length, a consistent predictor of preterm delivery; or by decreasing the mechanical support of the cervix, as increasing amounts of stromal tissue are excised. LEEP also results in the removal and permanent destruction of the mucus producing cells, which are most highly concentrated in the lower portion of the endocervical canal. This loss of glandular tissue may alter the composition and production of cervical secretions, and impact the antimicrobial, immunologic, and inflammatory milieu of the lower and upper genital tract. Thus, we propose to conduct a longitudinal clinic-based study of a cohort of non-pregnant women of reproductive age (18-35 yrs) requiring LEEP. Subjects will undergo ophthalimic sponge collection of cervical secretions and 3D transvaginal ultrasonography at the time of LEEP (n=50) and again six months later. Our first specific aim will determine if treatment by LEEP results in altered cervical length and volume. The fresh gross specimen will be measured with calipers to determine vertical cone height. Total tissue volume will be assessed by water displacement. Statistical analyses will determine if these pathological parameters correlate with changes in cervical length and volume following surgical healing. Our second aim will determine if LEEP alters the immunologic and antimicrobial biomarker profiles of cervical secretions using multiplex panel measurements of 22 analyses. For this aim, untreated women (n=50) who had a colposcopy with punch biopsy will be recruited from the same clinic to serve as a comparison group. These subjects should be similar to those requiring LEEP because they followed the same referral pathway for assessment of CIN. Changes in biomarker concentrations will be analyzed to determine associations with pathological correlates, while controlling for potential confounders. Paired statistical techniques, simple and multivariate linear regression, ANOVA, two-way repeated measures ANOVA, and ANCOVA will be used. [unreadable] [unreadable] [unreadable] [unreadable] [unreadable]